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1.
J Sep Sci ; 44(21): 3924-3932, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34459118

RESUMEN

ß-Sitosterol is a major bioactive constituent in plants with potent anticancer effects against many human cancer cells, but its bioavailability and therapeutic efficacy are limited by its poor solubility in water. In this study, C6 -imidazole chitosan, C6 -1-methylimidazole chitosan, C6 -1-ethylimidazole chitosan, C6 -1-vinylimidazole chitosan, C6 -1-allylimidazole chitosan, and C6 -1-butylimidazole chitosan were prepared to extract ß-sitosterol from edible oil samples via ultrasonic-assisted solid liquid extraction. The structures and properties of the newly synthesized products were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, and elemental analysis. The extraction abilities of the derivatives were tested in the experiment with high-performance liquid chromatography (limit of detection 0.21 µg/g and limit of quantification 0.67 µg/g), and the % relative standard deviation (<3.25%) and recovery values of the prepared chitosan derivatives toward ß-sitosterol (average: 100.20%) were acceptable. The spiked interday and intraday recoveries of ß-sitosterol were 102.60 ± 2.78 and 103.90 ± 3.04%, respectively. The actual amounts of ß-sitosterol extracted from three real samples using C6 -imidazole chitosan according to the solid phase extraction method were 3302.40, 901.70, and 2045.60 mg/kg for corn oil, olive oil, and pea oil, respectively.


Asunto(s)
Quitosano/química , Imidazoles/química , Aceites de Plantas/química , Sitoesteroles/aislamiento & purificación , Extracción en Fase Sólida/métodos , Cromatografía Líquida de Alta Presión , Límite de Detección , Modelos Lineales , Microondas , Reproducibilidad de los Resultados , Sitoesteroles/análisis , Sitoesteroles/química
2.
Int J Mol Sci ; 22(24)2021 Dec 08.
Artículo en Inglés | MEDLINE | ID: mdl-34948003

RESUMEN

Plant sterols (PSs) cannot be synthesized in mammals and are exclusively diet-derived. PSs cross the blood-brain barrier and may have anti-neuroinflammatory effects. Obesity is linked to lower intestinal uptake and blood levels of PSs, but its effects in terms of neuroinflammation-if any-remain unknown. We investigated the effect of high-fat diet-induced obesity on PSs in the brain and the effects of the PSs campesterol and ß-sitosterol on in vitro microglia activation. Sterols (cholesterol, precursors, PSs) and polyunsaturated fatty acid-derived lipid mediators were measured in the food, blood, liver and brain of C57BL/6J mice. Under a PSs-poor high-fat diet, PSs levels decreased in the blood, liver and brain (>50%). This effect was reversible after 2 weeks upon changing back to a chow diet. Inflammatory thromboxane B2 and prostaglandin D2 were inversely correlated to campesterol and ß-sitosterol levels in all brain regions. PSs content was determined post mortem in human cortex samples as well. In vitro, PSs accumulate in lipid rafts isolated from SIM-A9 microglia cell membranes. In summary, PSs levels in the blood, liver and brain were associated directly with PSs food content and inversely with BMI. PSs dampen pro-inflammatory lipid mediators in the brain. The identification of PSs in the human cortex in comparable concentration ranges implies the relevance of our findings for humans.


Asunto(s)
Dieta Alta en Grasa/efectos adversos , Ácidos Grasos Insaturados/análisis , Lipidómica/métodos , Microglía/citología , Enfermedades Neuroinflamatorias/metabolismo , Obesidad/metabolismo , Fitosteroles/análisis , Alimentación Animal , Animales , Células Cultivadas , Colesterol/análogos & derivados , Colesterol/análisis , Cromatografía Liquida , Modelos Animales de Enfermedad , Humanos , Hígado/química , Masculino , Ratones , Ratones Endogámicos C57BL , Microglía/efectos de los fármacos , Microglía/metabolismo , Enfermedades Neuroinflamatorias/inducido químicamente , Obesidad/inducido químicamente , Fitosteroles/sangre , Sitoesteroles/análisis , Espectrometría de Masas en Tándem
3.
Molecules ; 26(16)2021 Aug 12.
Artículo en Inglés | MEDLINE | ID: mdl-34443479

RESUMEN

Cannabis sativa L. has been used for a long time to obtain food, fiber, and as a medicinal and psychoactive plant. Today, the nutraceutical potential of C.sativa is being increasingly reappraised; however, C. sativa roots remain poorly studied, despite citations in the scientific literature. In this direction, we identified and quantified the presence of valuable bioactives (namely, ß-sitosterol, stigmasterol, campesterol, friedelin, and epi-friedelanol) in the root extracts of C. sativa, a finding which might pave the way to the exploitation of the therapeutic potential of all parts of the C. sativa plant. To facilitate root harvesting and processing, aeroponic (AP) and aeroponic-elicited cultures (AEP) were established and compared to soil-cultivated plants (SP). Interestingly, considerably increased plant growth-particularly of the roots-and a significant increase (up to 20-fold in the case of ß-sitosterol) in the total content of the aforementioned roots' bioactive molecules were observed in AP and AEP. In conclusion, aeroponics, an easy, standardized, contaminant-free cultivation technique, facilitates the harvesting/processing of roots along with a greater production of their secondary bioactive metabolites, which could be utilized in the formulation of health-promoting and health-care products.


Asunto(s)
Cannabis/química , Cannabis/crecimiento & desarrollo , Hidroponía , Colesterol/análogos & derivados , Colesterol/análisis , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análisis , Fitosteroles/análisis , Extractos Vegetales/química , Raíces de Plantas/química , Raíces de Plantas/crecimiento & desarrollo , Sitoesteroles/análisis , Estigmasterol/análisis , Triterpenos/análisis
4.
Molecules ; 26(4)2021 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-33672019

RESUMEN

Putrajeevak (Putranjiva roxburghii Wall.; synonym Drypetes roxburghii (Wall.) Hurus) seeds have been used since ancient times in the treatment of infertility in the Ayurvedic system of medicine in India. In this study, the oil component of Putrajeevak seeds (PJSO) was extracted using the supercritical fluid extraction (SCFE) method using liquid CO2 and the constituents were analyzed using gas chromatography-flame ionized detectorand high-performance thin-layer chromatography. PJSO contained trace amounts of ß-sitosterol with oleic and linoleic acids as the major fatty acid constituents. Male and female zebrafish were mutagenized with N-ethyl-N-nitrosourea (ENU) and fish that produced less than 20 viable embryos were selected for the study. SCFE oil extracts from the P. roxburghii seeds were used in this study to reverse fertility impairment. The mutant fish were fed with PJSO for a period of 14 days and the rates of fertility, conception, and fecundity were determined with wild-type healthy fish as a breeding partner. Treatment with PJSO increased the ovarian follicle count as well as the number of mature eggs, while reducing the number of ovarian cysts. Sperm count as well as sperm motility were greatly enhanced in the ENU-mutagenized male zebrafish when treated with PJSO. The results obtained in this study demonstrate the effectiveness of P. roxburghii seed oil in reversing impaired fertility in both male and female zebrafish models.


Asunto(s)
Cromatografía con Fluido Supercrítico/métodos , Euphorbiaceae/química , Fertilidad/fisiología , Semillas/química , Pez Cebra/fisiología , Animales , Modelos Animales de Enfermedad , Ácidos Grasos/análisis , Ácidos Grasos/química , Femenino , Fertilidad/efectos de los fármacos , Inflamación/tratamiento farmacológico , Inflamación/patología , Masculino , Ovario/fisiología , Óvulo/fisiología , Pelvis/patología , Aceites de Plantas/química , Aceites de Plantas/farmacología , Aceites de Plantas/uso terapéutico , Sitoesteroles/análisis , Sitoesteroles/química , Testículo/efectos de los fármacos , Testículo/patología
5.
Anal Chem ; 92(13): 8628-8637, 2020 07 07.
Artículo en Inglés | MEDLINE | ID: mdl-32510944

RESUMEN

Mass spectrometry (MS) in hyphenated techniques is widely accepted as the gold standard quantitative tool in life sciences. However, MS possesses intrinsic analytical capabilities that allow it to be a stand-alone quantitative technique, particularly with current technological advancements. MS has a great potential for simplifying quantitative analysis without the need for tedious chromatographic separation. Its selectivity relies on multistage MS analysis (MSn), including tandem mass spectrometry (MS/MS), as well as the ever-growing advancements of high-resolution MS instruments. This perspective describes various analytical platforms that utilize MS as a stand-alone quantitative technique, namely, flow injection analysis (FIA), matrix assisted laser desorption ionization (MALDI), including MALDI-MS imaging and ion mobility, particularly high-field asymmetric waveform ion mobility spectrometry (FAIMS). When MS alone is not capable of providing reliable quantitative data, instead of conventional liquid chromatography (LC)-MS, the use of a guard column (i.e., fast chromatography) may be sufficient for quantification. Although the omission of chromatographic separation simplifies the analytical process, extra procedures may be needed during sample preparation and clean-up to address the issue of matrix effects. The discussion of this manuscript focuses on key parameters underlying the uniqueness of each technique for its application in quantitative analysis without the need for a chromatographic separation. In addition, the potential for each analytical strategy and its challenges are discussed as well as improvements needed to render them as mainstream quantitative analytical tools. Overcoming the hurdles for fully validating a quantitative method will allow MS alone to eventually become an indispensable quantitative tool for clinical and toxicological studies.


Asunto(s)
Espectrometría de Masas/métodos , Colesterol/análogos & derivados , Colesterol/análisis , Cromatografía Líquida de Alta Presión , Análisis de Inyección de Flujo , Espectrometría de Movilidad Iónica , Límite de Detección , Fitosteroles/análisis , Sitoesteroles/análisis , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
6.
Acta Pharmacol Sin ; 41(9): 1178-1196, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32504068

RESUMEN

ß-Sitosterol (24-ethyl-5-cholestene-3-ol) is a common phytosterol Chinese medical plants that has been shown to possess antioxidant and anti-inflammatory activity. In this study we investigated the effects of ß-sitosterol on influenza virus-induced inflammation and acute lung injury and the molecular mechanisms. We demonstrate that ß-sitosterol (150-450 µg/mL) dose-dependently suppresses inflammatory response through NF-κB and p38 mitogen-activated protein kinase (MAPK) signaling in influenza A virus (IAV)-infected cells, which was accompanied by decreased induction of interferons (IFNs) (including Type I and III IFN). Furthermore, we revealed that the anti-inflammatory effect of ß-sitosterol resulted from its inhibitory effect on retinoic acid-inducible gene I (RIG-I) signaling, led to decreased STAT1 signaling, thus affecting the transcriptional activity of ISGF3 (interferon-stimulated gene factor 3) complexes and resulting in abrogation of the IAV-induced proinflammatory amplification effect in IFN-sensitized cells. Moreover, ß-sitosterol treatment attenuated RIG-I-mediated apoptotic injury of alveolar epithelial cells (AEC) via downregulation of pro-apoptotic factors. In a mouse model of influenza, pre-administration of ß-sitosterol (50, 200 mg·kg-1·d-1, i.g., for 2 days) dose-dependently ameliorated IAV-mediated recruitment of pathogenic cytotoxic T cells and immune dysregulation. In addition, pre-administration of ß-sitosterol protected mice from lethal IAV infection. Our data suggest that ß-sitosterol blocks the immune response mediated by RIG-I signaling and deleterious IFN production, providing a potential benefit for the treatment of influenza.


Asunto(s)
Lesión Pulmonar Aguda/tratamiento farmacológico , Antivirales/uso terapéutico , Proteína 58 DEAD Box/metabolismo , Inflamación/tratamiento farmacológico , Transducción de Señal/efectos de los fármacos , Sitoesteroles/uso terapéutico , Células A549 , Lesión Pulmonar Aguda/patología , Lesión Pulmonar Aguda/virología , Animales , Antivirales/análisis , Apoptosis/efectos de los fármacos , Perros , Femenino , Células HEK293 , Humanos , Inflamación/patología , Inflamación/virología , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Interferón Tipo I/metabolismo , Interferones/metabolismo , Pulmón/patología , Células de Riñón Canino Madin Darby , Ratones Endogámicos BALB C , Plantas/química , Factor de Transcripción STAT1/metabolismo , Sitoesteroles/análisis , Interferón lambda
7.
J Sep Sci ; 43(2): 418-437, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31650695

RESUMEN

A rapid and sensitive ultra high performance liquid chromatography with tandem mass spectrometry method was established and validated for simultaneous determination of thirteen bioactive components (gallic acid, protocatechuic acid, puerarin, p-hydroxycinnamic acid, daidzin, ononin, daidzein, naringenin, genistein, apigenin, formononetin, biochanin A, and ß-sitosterol) of Radix Puerariae extract in rat plasma and tissues. The plasma and tissues samples were pretreated by protein precipitation extraction, and umbelliferone and rutin were used as internal standards. Sample separation was performed on a ZORBAX RRHD Eclipse plus C18 column (2.1 mm × 50 mm, 1.8 µm, Agilent) with a mobile phase consisting of methanol-water (containing 0.1% formic acid). The mass spectrometry analysis was conducted in positive and negative ionization modes with multiple reaction monitoring. The lower limit of quantitation range for the 13 analytes was 0.2-35 ng/mL. The intra- and inter-day precision of all the analytes were less than 10.92%, with an accuracy ranging from -13.10 to 11.96%. Both the recovery and matrix effect were within acceptable limits. This method was successfully applied to pharmacokinetic and tissue distribution study of the 13 bioactive components in rats after oral administration of R. Puerariae extract.


Asunto(s)
Apigenina/farmacocinética , Medicamentos Herbarios Chinos/farmacocinética , Genisteína/farmacocinética , Isoflavonas/farmacocinética , Pueraria/química , Sitoesteroles/farmacocinética , Administración Oral , Animales , Apigenina/administración & dosificación , Apigenina/análisis , Cromatografía Líquida de Alta Presión , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/análisis , Genisteína/administración & dosificación , Genisteína/análisis , Isoflavonas/administración & dosificación , Isoflavonas/análisis , Estructura Molecular , Ratas , Ratas Sprague-Dawley , Sitoesteroles/administración & dosificación , Sitoesteroles/análisis , Espectrometría de Masas en Tándem , Distribución Tisular
8.
Molecules ; 25(9)2020 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-32392823

RESUMEN

Brevicoryne brassicae is a problematic pest in cabbage and other field crops. Synthetic pesticides are used to control this pest, but they are injurious for human health and the environment. The present study aimed to purify and identify the active compounds from Citrullus colocynthis leaves with an appraisal of their efficacy against B. brassicae. Separation and purification were performed via different chromatographic techniques. Molecular analysis and chemical structures were recognized by mass spectrum (MS) and nuclear magnetic resonance (NMR), respectively. Moreover, in vitro and in vivo aphicidal activity was assessed using various concentrations, i.e., 6.25, 12.5, 25 and 50 µg/mL at 12, 24, 48 and 72 h exposure. The outcome shows that mass spectrum analyses of the purified compounds suggested the molecular formulae are C30H50O and C29H50O, C29H48O. The compounds were characterized as fernenol and a mixture of spinasterol, 22,23-dihydrospinasterol by 1H-NMR and 13C-NMR spectrum analysis. The toxicity results showed that the mixture of spinasterol and 22,23-dihydrospinasterol showed LC50 values of 32.36, 44.49 and 37.50 µg/mL by contact, residual and greenhouse assay at 72 h exposure, respectively. In contrast, fernenol recorded LC50 values as 47.99, 57.46 and 58.67 µg/mL, respectively. On the other hand, spinasterol, 22,23-dihydrospinasterol showed the highest mortality, i.e., 66.67%, 53.33% and 60% while, 30%, 23.33% and 25% mortality was recorded by fernenol after 72 h at 50 µg/mL by contact, residual and greenhouse assay, respectively. This study suggests that spinasterol, 22,23-dihydrospinasterol are more effective against B. brassicae which may be introduced as an effective and suitable substitute of synthetic chemical pesticides.


Asunto(s)
Áfidos/efectos de los fármacos , Citrullus colocynthis/química , Insecticidas/toxicidad , Hojas de la Planta/química , Sitoesteroles/toxicidad , Estigmasterol/análogos & derivados , Triterpenos/toxicidad , Animales , Insecticidas/análisis , Insecticidas/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/toxicidad , Sitoesteroles/análisis , Sitoesteroles/química , Sitoesteroles/aislamiento & purificación , Estigmasterol/análisis , Estigmasterol/química , Estigmasterol/aislamiento & purificación , Estigmasterol/toxicidad , Triterpenos/análisis , Triterpenos/química , Triterpenos/aislamiento & purificación
9.
J Dairy Sci ; 102(9): 7723-7733, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31255261

RESUMEN

Milk fat adulteration is a common issue in Central Asia. To assess the current situation in the commercial milk market, 17 milk samples were checked for fatty acid (FA) and sterol profiles to detect potential adulteration using multivariate analysis. Analysis of FA and sterols was performed using gas chromatography with flame ionization detection and gas chromatography with mass-spectrometric detection, respectively. Cluster analysis of FA profiles revealed 3 types of milk samples: (1) samples containing a higher proportion of short-chain FA, (2) samples containing a higher proportion of long-chain FA, and (3) samples with significant amounts of C18 FA. Analysis of sterols showed that samples included (1) milk fat containing 100% cholesterol, sometimes with traces of phytosterols, (2) milk fat with high proportions of ß-sitosterol and campesterol, and (3) milk fat containing high proportions of brassicasterol. We found significant relationships between FA profiles and sterol profiles. The profiles were compared with vegetable oil patterns reported in the literature. More than 50% of the samples appeared to be counterfeited. We conclude that identification of adulteration in milk can be based solely on determination of sterol patterns.


Asunto(s)
Ácidos Grasos/análisis , Contaminación de Alimentos/análisis , Leche/química , Esteroles/análisis , Animales , Colestadienoles/análisis , Colesterol/análogos & derivados , Colesterol/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Fitosteroles/análisis , Aceites de Plantas/análisis , Sitoesteroles/análisis
10.
BMC Complement Altern Med ; 19(1): 154, 2019 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-31269948

RESUMEN

BACKGROUND: Of over 35 Saudi plants traditionally used to treat liver disorders, majority still lack scientific validations. We therefore, evaluated the anti-oxidative, anti-apoptotic and hepatoprotective potential of Solanum surattense leaves total ethanol-extract (SSEE). METHODS: The cytoprotective (4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide/ MTT assay) and anti-apoptotic (caspase-3/7) potential of SSEE (25-200 µg/mL) were assessed in cultured HepG2 cells against dichlorofluorescein (DCFH)-induced toxicity. The hepatoprotective salutation of SSEE (100 and 200 mg/kg.bw/day) in carbon tetrachloride (CCl4)-intoxicated rats was evaluated by serum biochemistry and histopathology. The anti-oxidative activity of SSEE (31.25-500 µg/mL) was tested by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging and linoleic acid bleaching assays. Also, SSEE was subjected to qualitative phytochemical analysis, and standardized by validated high-performance liquid chromatography (HPTLC). RESULTS: SSEE at doses 50, 100 and 200 µg/mL showed HepG2 cell proliferative and protective potential by about 61.0, 67.2 and 95%, respectively through inhibition of caspase-3/7 against DCFH-toxicity. In CCl4-injured rats, SSEE (200 mg/kg) significantly (P < 0.001) normalized serum transaminases, alkaline phosphatase, bilirubin, cholesterol, triglycerides, and total protein, including tissue malondialdehyde and nonprotein sulfhydryls levels, supported by the liver histopathology. SSEE further showed strong in vitro anti-oxidative and anti-lipid peroxidative activities, evidenced by the presence of alkaloids, flavonoids, tannins, sterols and saponins. Identification of ß-sitosterol (3.46 µg/mg) strongly supported the anti-oxidative and hepatoprotective salutation of SSEE. CONCLUSION: Our findings suggest the therapeutic potential of S. surattense against chemical-induced oxidative stress and liver damage. However, isolation of the active principles and elucidation of mechanism of action remain to be addressed.


Asunto(s)
Antioxidantes/análisis , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Hígado/efectos de los fármacos , Extractos Vegetales/farmacología , Solanum/química , Animales , Apoptosis/efectos de los fármacos , Intoxicación por Tetracloruro de Carbono/tratamiento farmacológico , Evaluación Preclínica de Medicamentos , Células Hep G2 , Humanos , Extractos Vegetales/química , Extractos Vegetales/uso terapéutico , Ratas Wistar , Sitoesteroles/análisis
11.
BMC Complement Altern Med ; 19(1): 113, 2019 Jun 03.
Artículo en Inglés | MEDLINE | ID: mdl-31159797

RESUMEN

BACKGROUND: Embryonic neural stem cells (eNSCs) are immature precursors of the central nervous system (CNS), with self-renewal and multipotential differentiation capacities. These are regulated by endogenous and exogenous factors such as alpha-linolenic acid (ALA), a plant-based essential omega-3 polyunsaturated fatty acid. METHODS: In this study, we investigated the effects of various concentrations of Alyssum homolocarpum seed oil (AHSO), containing natural ALA, stearic acid (SA), myristic acid (MA), and ß-sitosterol, on proliferation and differentiation of eNSCs, in comparison to controls and to synthetic pure ALA. RESULTS: Treatment with natural AHSO (25 to 75 µM), similar to synthetic ALA, caused a significant ~ 2-fold increase in eNCSs viability, in comparison to controls. To confirm this proliferative activity, treatment of NSCs with 50 or 75 µM AHSO resulted in a significant increase in mRNA levels of notch1, hes-1 and Ki-67and NICD protein expression, in comparison to controls. Moreover, AHSO administration significantly increased the differentiation of eNSCs toward astrocytes (GFAP+) and oligodendrocytes (MBP+) in a dose dependent manner and was more potent than ALA, at similar concentrations, in comparison to controls. Indeed, only high concentrations of 100 µM AHSO, but not ALA, caused a significant increase in the frequency of neurons (ß-III Tubulin+). CONCLUSION: Our data demonstrated that AHSO, a rich source of ALA containing also other beneficial fatty acids, increased the proliferation and stimulated the differentiation of eNSCs. We suggest that AHSO's effects are caused by ß-sitosterol, SA and MA, present within this oil. AHSO could be used in diet to prevent neurodevelopmental syndromes, cognitive decline during aging, and various psychiatric disorders.


Asunto(s)
Brassicaceae/química , Células-Madre Neurales/efectos de los fármacos , Neurogénesis/efectos de los fármacos , Aceites de Plantas/farmacología , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Antígeno Ki-67/metabolismo , Ratones , Ácido Mirístico/análisis , Células-Madre Neurales/metabolismo , Aceites de Plantas/química , Semillas/química , Sitoesteroles/análisis , Ácidos Esteáricos/análisis , Ácido alfa-Linolénico/análisis
12.
Molecules ; 24(21)2019 Oct 31.
Artículo en Inglés | MEDLINE | ID: mdl-31683500

RESUMEN

ß-sitosterol ß-d-glucoside (BSSG) was extracted from "piña" of the Agave angustifolia Haw plant by microwave-assisted extraction (MAE) with a KOH solution such as a catalyst and a conventional maceration method to determine the best technique in terms of yield, extraction time, and recovery. The quantification and characterization of BSSG were done by high-performance thin layer chromatography (HPTLC), Fourier-transform infrared spectroscopy (FT-IR), and high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). With an extraction time of 5 s by MAE, a higher amount of BSSG (124.76 mg of ß-sitosterol ß-d-glucoside/g dry weight of the extract) than those for MAE extraction times of 10 and 15 s (106.19 and 103.97 mg/g dry weight respectively) was shown. The quantification of BSSG in the extract obtained by 48 h of conventional maceration was about 4-5 times less (26.67 mg/g dry weight of the extract) than the yields reached by the MAE treatments. MAE achieved the highest amount of BSSG, in the shortest extraction time while preserving the integrity of the compound's structure.


Asunto(s)
Agave/química , Etanol/química , Microondas , Extractos Vegetales/química , Sitoesteroles/análisis , Cromatografía en Capa Delgada , Estándares de Referencia , Sitoesteroles/química , Soluciones , Espectrometría de Masa por Ionización de Electrospray , Espectroscopía Infrarroja por Transformada de Fourier
13.
Anal Chem ; 90(14): 8487-8494, 2018 07 17.
Artículo en Inglés | MEDLINE | ID: mdl-29920210

RESUMEN

There has been an increasing interest during recent years in the role of the gut microbiome on health and disease. Therefore, metabolites in human feces related to microbial activity are attractive surrogate marker to track changes of microbiota induced by diet or disease. Such markers include 5α/ß-stanols as microbiome-derived metabolites of sterols. Currently, reliable, robust, and fast methods to quantify fecal sterols and their related metabolites are missing. We developed a liquid chromatography-high-resolution mass spectrometry (LC-MS/HRMS) method for the quantification of sterols and their 5α/ß-stanols in human fecal samples. Fecal sterols were extracted and derivatized to N, N-dimethylglycine esters. The method includes cholesterol, coprostanol, cholestanol and sitosterol, 5α/ß-sitostanol, campesterol and 5α/ß-campestanol. Application of a biphenyl column permits separation of isomeric 5α- and 5ß-stanols. Sterols are detected in parallel reaction monitoring (PRM) mode and stanols in full scan mode. HRMS allows differentiation of isobaric ß-stanols and the [M + 2] isotope peak of the coeluting sterol. Performance characteristics meet the criteria recommended by Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines. Analysis of fecal samples from healthy volunteers revealed high interindividual variability of sterol and stanol fractions. Interestingly, cholesterol and sitosterol showed similar fractions of mainly 5ß-stanols. In contrast, campesterol is substantially converted to 5α-campestanol and might be a poorer substrate for bacterial metabolism. Robust and fast quantification of fecal sterols and their related stanols by LC-MS/HRMS offers great potential to find novel microbiome-related biomarker in large-scale studies.


Asunto(s)
Heces/química , Microbioma Gastrointestinal , Esteroles/análisis , Espectrometría de Masas en Tándem/métodos , Colesterol/análogos & derivados , Colesterol/análisis , Cromatografía Liquida/economía , Cromatografía Liquida/métodos , Heces/microbiología , Humanos , Límite de Detección , Fitosteroles/análisis , Sitoesteroles/análisis , Espectrometría de Masas en Tándem/economía
14.
Biochem J ; 474(17): 2993-3010, 2017 08 17.
Artículo en Inglés | MEDLINE | ID: mdl-28739601

RESUMEN

Arthritis is a chronic inflammatory disease which reduces the life quality of affected individuals. Therapeutic tools used for treating inflammatory pain are associated with several undesirable effects. Buddleja thyrsoides Lam., known as 'Barbasco' or 'Cambara', is mostly used in several disorders and possesses antirheumatic, anti-inflammatory, and analgesic properties. Here, we investigated the antinociceptive and anti-inflammatory effects of the B. thyrsoides crude extract applied orally and topically in acute pain models and an arthritic pain model induced by complete Freund's adjuvant (CFA) paw injection in male mice (25-30 g). The high-performance liquid chromatography (HPLC) of the B. thyrsoides extract crude revealed the presence of the lupeol, stigmasterol, and ß-sitosterol. The stability study of the B. thyrsoides gel did not show relevant changes at low temperatures. The oral treatment with the B. thrysoides extract prevented the capsaicin-induced spontaneous nociception and the acetic acid-induced abdominal writhing, but did not alter the thermal threshold in the tail immersion test. The B. thyrsoides antinociceptive effect was not reversed by naloxone in the capsaicin test. The B. thyrsoides oral or topical treatment reversed the CFA-induced mechanical allodynia and thermal hyperalgesia with maximum inhibition (Imax) of 69 ± 6 and 68 ± 5% as well as 78 ± 15 and 87 ± 12%, respectively. Moreover, the topical but not oral treatment inhibited the CFA-induced cell infiltration, but did not reduce the paw edema significantly. The oral treatment with B. thyrsoides did not cause adverse effects. These findings suggest that the oral or topical treatment with B. thyrsoides presents antinociceptive actions in an arthritic pain model without causing adverse effects.


Asunto(s)
Analgésicos no Narcóticos/uso terapéutico , Antiinflamatorios no Esteroideos/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Buddleja/química , Extractos Vegetales/uso terapéutico , Hojas de la Planta/química , Dolor Agudo/tratamiento farmacológico , Administración Cutánea , Administración Oral , Analgésicos no Narcóticos/administración & dosificación , Analgésicos no Narcóticos/efectos adversos , Analgésicos no Narcóticos/química , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/química , Brasil , Buddleja/crecimiento & desarrollo , Estabilidad de Medicamentos , Almacenaje de Medicamentos , Etnofarmacología , Geles , Calor/efectos adversos , Masculino , Ratones , Triterpenos Pentacíclicos/administración & dosificación , Triterpenos Pentacíclicos/efectos adversos , Triterpenos Pentacíclicos/análisis , Triterpenos Pentacíclicos/uso terapéutico , Extractos Vegetales/administración & dosificación , Extractos Vegetales/efectos adversos , Extractos Vegetales/química , Hojas de la Planta/crecimiento & desarrollo , Sitoesteroles/administración & dosificación , Sitoesteroles/efectos adversos , Sitoesteroles/análisis , Sitoesteroles/uso terapéutico , Estigmasterol/administración & dosificación , Estigmasterol/efectos adversos , Estigmasterol/análisis , Estigmasterol/uso terapéutico , Viscosidad
15.
BMC Complement Altern Med ; 18(1): 233, 2018 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-30086745

RESUMEN

BACKGROUND: Bombax ceiba is used traditionally to treat bone disorders, rheumatism, and joint pain. The aim of the study is to carry out osteogenic activity in-vitro and anti-osteoporotic activity in-vivo of stem bark of B. ceiba in surgical ovariectomy model in female rats. METHODS: Plant drug: B. ceiba stem bark was extracted with solvents petroleum ether and methanol using Soxhlet extraction. In-vitro osteoblastic proliferation study was performed using UMR-106 cell lines. Both the extracts were undergone to acute toxicity study as per OECD423 guidelines. Female Wistar albino rats 180-240 g were used (n = 6). Surgical ovariectomy was performed under anesthesia to induce bone porosity and loss in all animals except normal control and sham control. Each extract was administered at two dose level: 100 and 200 mg/kg and the standard Raloxifene was given at 1 mg/kg orally for 28 days. The phytochemical study of both the extracts was performed using HPLC and HPTLC. RESULTS: A significant osteoblast cell proliferation and alkaline phosphatase activity were observed with B. ceiba extracts in UMR-106 cell lines. Surgical removal of ovaries produced significant (p < 0.05) decline in bone mineral density, bone breaking strength, serum ALP, calcium, phosphorus, and estradiol level and marked bone tissue destruction in histology. Administration of petroleum ether and methanolic extract for 28 days significantly (p < 0.05) ameliorated the consequences of ovariectomy induced bone porosity and restored the normal architecture of bone, as compared to OVX control. The phytochemical screening of both the extracts were also carried out. The quantification of phytoconstituents showed the presence of ß-sitosterol and lupeol in petroleum ether extract, whereas the lupeol is also quantified in the methanolic extract. The presence of gallic acid was quantified in methanolic extract using HPLC. CONCLUSION: B. ceiba: stem bark ameliorated the state of bone fragility and fracture possibly due to estrogenic modulation, as also confirmed by in-vitro osteogenic activity which may be due to the presence of lupeol, gallic acid and ß-sitosterol constituents of the plant.


Asunto(s)
Bombax/química , Proliferación Celular/efectos de los fármacos , Ácido Gálico/farmacología , Osteoblastos/efectos de los fármacos , Osteoporosis/metabolismo , Triterpenos Pentacíclicos/farmacología , Sitoesteroles/farmacología , Animales , Peso Corporal/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Femenino , Ácido Gálico/análisis , Ovariectomía , Triterpenos Pentacíclicos/análisis , Extractos Vegetales/química , Extractos Vegetales/farmacología , Ratas , Ratas Wistar , Sitoesteroles/análisis
16.
Int J Food Sci Nutr ; 68(6): 695-703, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28276904

RESUMEN

Sterols are components present in the fat fraction of infant formulas (IFs). Their characterization is therefore of interest, though there are no official reference methods for their analysis in these matrices. AIM: To validate a gas chromatographic method with flame ionization detection for the determination of animal (cholesterol and desmosterol) and plant sterols (brassicasterol, campesterol, stigmasterol, ß-sitosterol and sitostanol) found in IFs. All correlation coefficients obtained for the calibration curves of sterols studied were >0.99. Limits of detection (<1 µg/100 mL) and quantification (<4 µg/100 mL) are suitable for sterols determination in IFs. The within-assay precision ranged from 1.6% to 8.8%, while the between-assay precision was <10% for most of sterols. Accuracy was satisfactory and was calculated by recovery assays (ranging 93-108%). The analytical parameters obtained showed the suitability of the proposed method for the determination of sterols in IFs.


Asunto(s)
Cromatografía de Gases , Fórmulas Infantiles/química , Fitosteroles/análisis , Calibración , Colestadienoles/análisis , Colesterol/análogos & derivados , Colesterol/análisis , Desmosterol/análisis , Ionización de Llama , Límite de Detección , Reproducibilidad de los Resultados , Sitoesteroles/análisis , Estigmasterol/análisis
17.
Int J Food Sci Nutr ; 68(1): 52-64, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27575665

RESUMEN

Chicories produce a wide range of vegetables with important nutritional value. We determined the variation of sterol, total polyphenol, nitrate contents and antioxidant capacity (SC, TPC, NC, AC) in endive leaves and stem-chicory novel vegetables, cultivated in two Italian regions. Within a given area, the SC was similar in smooth- and curly leafed endives (106.3-176.0 mg/kg FW); sitosterol and stigmasterol were major fractions (45-56 versus 38-43%). The stem SC was independent of landrace (101.5-118.6 mg/kg FW); sitosterol prevailed on stigmasterol and fucosterol (73-76 versus 12-14% versus 8-9%); the latter reached 15.7 mg/kg FW, conferring value as potential antidiabetes food. The planting site affected the AC and TPC of endives (893.1-1571.4 µmTE/100 g FW, 30.8-76.1 GAE100/g FW) and chicory stems (729.8-1152.5 µmTE/100 g FW; 56.2-124.4 GAE100/g FW), while the NC was recurrently below dangerous thresholds. PCA showed that environment was the major cause of variation, though it modestly affected these parameters.


Asunto(s)
Antioxidantes/análisis , Asteraceae/química , Cichorium intybus/química , Productos Agrícolas/química , Fitosteroles/análisis , Hojas de la Planta/química , Tallos de la Planta/química , Antioxidantes/química , Antioxidantes/metabolismo , Asteraceae/crecimiento & desarrollo , Asteraceae/metabolismo , Cichorium intybus/crecimiento & desarrollo , Cichorium intybus/metabolismo , Producción de Cultivos , Productos Agrícolas/crecimiento & desarrollo , Productos Agrícolas/metabolismo , Contaminación de Alimentos , Alimentos Funcionales/análisis , Humanos , Italia , Nitratos/análisis , Nitratos/química , Nitratos/metabolismo , Valor Nutritivo , Capacidad de Absorbancia de Radicales de Oxígeno , Fenoles/análisis , Fenoles/química , Fenoles/metabolismo , Fitosteroles/biosíntesis , Fitosteroles/química , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Tallos de la Planta/crecimiento & desarrollo , Tallos de la Planta/metabolismo , Análisis de Componente Principal , Sitoesteroles/análisis , Sitoesteroles/química , Sitoesteroles/metabolismo , Análisis Espacio-Temporal , Especificidad de la Especie , Estigmasterol/análogos & derivados , Estigmasterol/análisis , Estigmasterol/química , Estigmasterol/metabolismo
18.
Z Naturforsch C J Biosci ; 71(3-4): 65-71, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26959540

RESUMEN

A phytochemical study of Ficus thonningii has led to the isolation of two previously unreported compounds, thonningiiflavanonol A and thonningiiflavanonol B together with 16 known compounds: shuterin, naringenin, syringic acid, p-hydroxybenzoic acid, genistein, 5,7,3',4',5'-pentahydroxyflavanone, luteolin, methylparaben, aromadendrin, garbanzol, dihydroquercetin, 5,7,3'-trihydroxyflavanone, ß-sitosterol, sitosterolglucoside, lupeol acetate, and taraxerol. Their structures were elucidated on the basis of spectroscopic data. The new compounds and extracts displayed potent antioxidant activity.


Asunto(s)
Ficus/química , Flavonoides/análisis , Corteza de la Planta/química , Extractos Vegetales/química , Raíces de Plantas/química , Tallos de la Planta/química , Antioxidantes/análisis , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Flavanonas/análisis , Flavanonas/química , Flavanonas/aislamiento & purificación , Flavonoides/química , Flavonoides/aislamiento & purificación , Ácido Gálico/análogos & derivados , Ácido Gálico/análisis , Ácido Gálico/química , Ácido Gálico/aislamiento & purificación , Genisteína/análisis , Genisteína/química , Genisteína/aislamiento & purificación , Luteolina/análisis , Luteolina/química , Luteolina/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Estructura Molecular , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/análisis , Ácido Oleanólico/química , Ácido Oleanólico/aislamiento & purificación , Parabenos/análisis , Parabenos/química , Parabenos/aislamiento & purificación , Quercetina/análogos & derivados , Quercetina/análisis , Quercetina/química , Quercetina/aislamiento & purificación , Sitoesteroles/análisis , Sitoesteroles/química , Sitoesteroles/aislamiento & purificación
19.
J Sci Food Agric ; 96(12): 4143-50, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26762864

RESUMEN

BACKGROUND: In olive oil, sterols constitute the majority of the unsaponifiable fraction. In recent years there has been increased interest in the sterols of olive oil for their health benefits and their importance to virgin olive oil (VOO) quality regulation. RESULTS: Forty-three olive (Olea europaea L.) cultivars from the World Olive Germplasm Bank, IFAPA Centro 'Alameda de Obispo', Cordoba, Spain were studied for their oil sterol composition and total content. The main sterols found in olive oil were ß-sitosterol, Δ(5) -avenasterol, campesterol and stigmasterol, most of them showing high variability. Most cultivars showed total sterol contents within the limits established by EU regulations, although 28% of VOOs analysed were outside the limits established for total content and/or for individual sterols. Over the group of cultivars, total sterol contents ranged from 855 to 2185 mg kg(-1) . CONCLUSION: The high variability observed was due to the genetic component, since other agronomic and technological factors were similar. Because of the high variability, the sterol fraction can be considered as a useful tool to characterize and discriminate monovarietal VOOs. The results can be useful for nutritionists for VOO inclusion in nutrition studies. Furthermore, the variability observed can be applied in olive breeding projects to select the parents of new olive cultivars with an improved sterol fraction. © 2016 Society of Chemical Industry.


Asunto(s)
Olea/química , Aceite de Oliva/química , Esteroles/análisis , Industria Química , Frutas/química , Genotipo , Olea/clasificación , Olea/genética , Fitomejoramiento , Banco de Semillas , Sitoesteroles/análisis , España , Especificidad de la Especie
20.
J Sci Food Agric ; 96(9): 3256-65, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26522347

RESUMEN

BACKGROUND: The by-products of seven different cereal grains were investigated as a source of extractable oil, rich in lipophilic bioactive compounds. RESULTS: Oil yields (g kg(-1) DW) recovered from cereal by-products were as follows: 189 (rice bran) > 112 (wheat germ) > 74 (corn bran) > 58 (oat bran) > 41 (buckwheat bran) > 39 (spelt bran) > 33 (wheat bran) > 27 (rye bran). The main fatty acids identified in the studied oil samples were palmitic acid (11.39-17.23%), oleic acid (11.76-42.73%), linoleic acid (35.54-62.65%) and α-linolenic acid (1.05-9.46%). The range of total tocochromanols and phytosterols in the obtained oils was 0.369-3.763 and 1.19-35.24 g kg(-1) of oil, respectively. The oils recovered from buckwheat and corn bran, and wheat germ were dominated by tocopherols (99.9, 84.2 and 96.5%, respectively), whereas the oat, rice, rye, spelt, wheat bran oils were rich in tocotrienols (73.9, 79.6, 78.1, 90.6 and 73.8%, respectively). The campesterol and ß-sitosterol constituted 10.1-32.5 and 30.4-63.7%, respectively, of total phytosterols contents identified in all of the studied samples. CONCLUSION: The present study demonstrated that oils recovered from the cereal by-products are richer sources of bioactive compounds, compared with traditional oils. © 2015 Society of Chemical Industry.


Asunto(s)
Grano Comestible/química , Aceites de Plantas/química , Colesterol/análogos & derivados , Colesterol/análisis , Ácidos Grasos/análisis , Fitosteroles/análisis , Extractos Vegetales/química , Semillas/química , Sitoesteroles/análisis , Escualeno/análisis , Tocoferoles/análisis , Tocotrienoles/análisis , Triticum/química
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